Talin and Signaling Through Integrins

Integrin adhesion receptors are essential for the development and functioning of multicellular animals. Integrins mediate cell adhesion to the extracellular matrix and to counter-receptors on adjacent cells, and the ability of integrins to bind extracellular ligands is regulated in response to intracellular signals that act on the short cytoplasmic tails of integrin subunits. Integrin activation, the rapid conversion of integrin receptors from low to high affinity, requires binding of talin to integrin β tails and, once bound, talin provides a connection from activated integrins to the actin cytoskeleton. A wide range of experimental approaches have contributed to the current understanding of the importance of talin in integrin signaling. Here, we describe two methods that have been central to our investigations of talin; a biochemical assay that has allowed characterization of interactions between integrin cytoplasmic tails and talin, and a fluorescent-activated cell-sorting procedure to assess integrin activation in cultured cells expressing talin domains, mutants, dominant negative constructs, or shRNA.